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目的研究昆布多糖硫酸酯(LAPS)对过氧化氢(H2O2)诱导乳鼠心肌细胞氧化损伤的保护作用。方法分次消化法分离原代培养的乳鼠心肌细胞,建立H2O2诱导心肌细胞损伤模型;四甲基偶氮唑盐(MTT)法检测不同浓度LAPS对心肌细胞的保护作用;试剂盒检测LAPS对细胞乳酸脱氢酶(LDH)、肌酸磷酸激酶(CK)、丙二醛(MDA)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)含量的影响;利用荧光探针二氯二氢荧光素-乙酰乙酸酯(DCFH-DA)检测细胞内活性氧(ROS)含量。结果100μmol/L的H2O2能明显造成心肌细胞损伤,LAPS能改善H2O2所造成的心肌细胞损伤;LAPS能够明显降低的过氧化损伤心肌细胞外液中LDH和CK含量、明显降低过氧化损伤心肌细胞内MDA和活性氧(ROS)的含量、明显提高过氧化损伤心肌细胞内SOD、GSH-Px和CAT的活力。结论 LAPS对心肌细胞的过氧化损伤有保护作用,与其降低或阻断脂质过氧化反应,降低细胞内ROS含量,进而阻断因ROS对细胞造成的损害,及时修复受损细胞有着密切关系。
Objective To investigate the protective effect of laminarin sulfate (LPS) on oxidative damage of neonatal rat cardiomyocytes induced by hydrogen peroxide (H2O2). Methods Primary cultured neonatal rat cardiomyocytes were isolated by graded digestion method to establish a model of cardiomyocyte injury induced by H2O2. MTT assay was used to detect the protective effects of different concentrations of LAPS on cardiomyocytes. (LDH), creatine phosphokinase (CK), malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase GSH-Px). The content of reactive oxygen species (ROS) in cells was detected by fluorescent probe dichlorodihydrofluorescein-acetoacetate (DCFH-DA). Results 100μmol / L H2O2 could significantly induce cardiomyocyte injury and LAPS could improve the damage of cardiomyocytes induced by H2O2. LAPS could significantly decrease the content of LDH and CK in pericardial extracellular fluid of peroxisome, MDA and reactive oxygen species (ROS), significantly increased the activity of SOD, GSH-Px and CAT in peroxisome damage myocardial cells. Conclusions LAPS has a protective effect on peroxidative injury of cardiomyocytes. It has a close relationship with reducing or blocking lipid peroxidation, decreasing intracellular ROS content, and then blocking the damage caused by ROS to cells and repairing injured cells promptly.